FIG 4.

UL148A requires a viral interaction partner external to the ULb′ region. (A to C) FLS1 HFFs (MICA*004/*009:01-*049) transduced with an EV (A), with UL148A (B), or with C-His UL148A (C) were either mock infected (black histograms) or VarS infected (red histograms). Cells were harvested 24 hpi, and MICA surface expression was assessed by flow cytometry. Gray-filled histograms represent isotype control staining of mock-infected cells, which produced similar results for all cells. (D and E) FLS1 HFFs (MICA*004/*009:01-*049) transduced with an EV or with C-His UL148A were either mock infected or infected with VarS or BAC2. (D) Cells were lysed 24 hpi, and Western blotting was performed using anti-MICA antibody for detection of MICA, anti-HIS antibody for detection of UL148A, and anti-vinculin antibody as a loading control. (E) Quantification of two independent experiments. MICA protein levels were quantified relative to the loading control (vinculin). RQ, relative quantification. Error bars represent SEM. Student's t test was performed to evaluate significance. *, P < 0.05. (F) FLS1 HFFs (MICA*004/*009:01-*049) transduced with an EV or with C-His UL148A were either mock infected or infected with mutant BAC2 ΔUL148A or BAC2. (F) Cells were lysed 24 hpi, and Western blotting was performed using anti-MICA antibody for detection of MICA, anti-HIS antibody for detection of UL148A, and anti-vinculin antibody as a loading control.